4afd4150ef5616ecb5fe48f288db02d6 intjoumolsci_a2016v17n4p552.pdf 5700cb8532f710d3a870f096f35d89bca55d8b5c intjoumolsci_a2016v17n4p552.pdf 572b6188bec060236b27585dafbcbb7d9de9ae15054195b48d39450267cfed1e intjoumolsci_a2016v17n4p552.pdf Title: Insights into the Antimicrobial Mechanism of Action of Human RNase6: Structural Determinants for Bacterial Cell Agglutination and Membrane Permeation Subject: Human Ribonuclease 6 is a secreted protein belonging to the ribonuclease A (RNaseA) superfamily, a vertebrate specific family suggested to arise with an ancestral host defense role. Tissue distribution analysis revealed its expression in innate cell types, showing abundance in monocytes and neutrophils. Recent evidence of induction of the protein expression by bacterial infection suggested an antipathogen function in vivo. In our laboratory, the antimicrobial properties of the protein have been evaluated against Gram-negative and Gram-positive species and its mechanism of action was characterized using a membrane model. Interestingly, our results indicate that RNase6, as previously reported for RNase3, is able to specifically agglutinate Gram-negative bacteria as a main trait of its antimicrobial activity. Moreover, a side by side comparative analysis with the RN6(1–45) derived peptide highlights that the antimicrobial activity is mostly retained at the protein N-terminus. Further work by site directed mutagenesis and structural analysis has identified two residues involved in the protein antimicrobial action (Trp1 and Ile13) that are essential for the cell agglutination properties. This is the first structure-functional characterization of RNase6 antimicrobial properties, supporting its contribution to the infection focus clearance. Keywords: RNases; host defence; antimicrobial peptides; cell agglutination; infectious diseases Author: David Pulido, Javier Arranz-Trullén, Guillem Prats-Ejarque, Diego Velázquez, Marc Torrent, Mohammed Moussaoui and Ester Boix Creator: LaTeX with hyperref package Producer: pdfTeX-1.40.15 CreationDate: Wed Apr 13 18:07:10 2016 ModDate: Wed Apr 13 18:07:10 2016 Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 19 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 3780815 bytes Optimized: no PDF version: 1.5 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- TCZUMN+URWPalladioL-Ital Type 1 Custom yes yes no 141 0 SDWVAE+URWPalladioL-Bold Type 1 Custom yes yes no 142 0 PTIHCV+URWPalladioL-Roma Type 1 Custom yes yes no 143 0 CKUPDE+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 147 0 IRWHZX+TimesNewRomanPSMT Type 1C WinAnsi yes yes no 148 0 UNSLQC+EURM10 Type 1 Builtin yes yes no 215 0 BMBFIK+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 220 0 BMBFJM+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 221 0 BMBFJO+PalatinoLinotype TrueType WinAnsi yes yes no 222 0 BMBFLP+TimesNewRoman TrueType WinAnsi yes yes no 223 0 FKPFBV+PazoMath-Italic Type 1 Builtin yes yes no 247 0 PZXOUX+TeX-matha9 Type 1 Builtin yes yes no 250 0 MKVSLO+TeX-matha8 Type 1 Builtin yes yes no 251 0 KCKEPA+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 268 0 KCKFAB+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 269 0 KCKFAD+PalatinoLinotype TrueType WinAnsi yes yes no 270 0 KCKFKB+TimesNewRoman TrueType WinAnsi yes yes no 271 0 VKMMUE+URWPalladioL-BoldItal Type 1 Custom yes yes no 295 0 KCKHGI+PalatinoLinotype,BoldItalic TrueType WinAnsi yes yes no 314 0 KCKEPA+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 315 0 KCKFAB+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 316 0 KCKFAD+PalatinoLinotype TrueType WinAnsi yes yes no 317 0 KCKFKB+TimesNewRoman TrueType WinAnsi yes yes no 318 0 KCKEPA+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 351 0 KCKFAB+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 352 0 KCKFAD+PalatinoLinotype TrueType WinAnsi yes yes no 353 0 KCKFKB+TimesNewRoman TrueType WinAnsi yes yes no 354 0 XNDZXZ+TeX-matha10 Type 1 Builtin yes yes no 384 0 KCKEPA+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 390 0 KCKFAB+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 391 0 KCKFAD+PalatinoLinotype TrueType WinAnsi yes yes no 392 0 KCKFKB+TimesNewRoman TrueType WinAnsi yes yes no 393 0 Jhove (Rel. 1.6, 2011-01-04) Date: 2016-11-22 02:33:49 CET RepresentationInformation: intjoumolsci_a2016v17n4p552.pdf ReportingModule: BYTESTREAM, Rel. 1.3 (2007-04-10) LastModified: 2016-11-21 11:31:29 CET Size: 3780815 Format: bytestream Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/octet-stream Checksum: b0c40298 Type: CRC32 Checksum: 4afd4150ef5616ecb5fe48f288db02d6 Type: MD5 Checksum: 5700cb8532f710d3a870f096f35d89bca55d8b5c Type: SHA-1